Clonal Evolution and Blast Crisis Correlate with Enhanced Proteolytic Activity of Separase in BCR-ABL b3a2 Fusion Type CML under Imatinib Therapy.

Unbalanced (major route) additional cytogenetic aberrations (ACA) in the diagnosis of chronic myeloid leukemia (CML) showed an increase in the risk of progression and shorter survival. In addition, the emerging ACA under imatinib treatment and clonal evolution regarded acceleration features and determine treatment failure in accordance with the European LeukemiaNet (ELN) recommendation.

On the basis of 1151 patients with chronic phase Philadelphia chromosome positive CML random-IV study, we examine recent events, ACA arising under imatinib treatment in connection with p210BCR-ABL breakpoint variants b2a2 and b3a2. We found the ACA preferential acquisition is not balanced in patients with vs b2a2 b3a2 fusion type (ratio: 6.3 vs. 1.6, p = 0.0246) concurring quickly advanced to the blast crisis for b3a2 patients (p = 0.0124).

ESPL1 / Separase, a cysteine ​​endopeptidase, is a key player in chromosome segregation during mitosis. Separase excess and / or hyperactivity have been reported from a variety of cancers and causes defective mitotic spindle, the chromosomes missegregation and aneuploidy. We investigated the influence of p210BCR-ABL breakpoint variants and treatment of imatinib on expression and proteolytic activity of Separase measured by specific fluorogenic assay in CML cell lines (b2a2: KCL-22, BV-173; b3a2: K562, LAMA-84). Despite the decline in protein content Separase up to 5.4-fold increase from Separase activity under imatinib treatment was observed exclusively in b3a2 but not in cell lines b2a2.

Mimic the effect of imatinib on BV-173 and LAMA-84 cells with silencing ESPL1 Separase stimulated proteolytic activity in cell lines of both b3a2 and b2a2. Our data indicate the presence of a feedback mechanism-related types of post- translational fusion Separase stimulates proteolytic activity after therapy-induced decrease in protein levels Separase. This can make b3a2 CML cells more susceptible to aneuploidy and clonal evolution from a common ancestor b2a2 and therefore can explain the results of cytogenetic patients with CML.

Separase Cleaves N-Tail of CENP-A CPAR Related Protein-1 in Meiosis I Metaphase-Anaphase transition in C. elegans.

Epigenetically defined centromere in most eukaryotes by the presence of centromeric chromatin containing histone H3 variant CENP-A. Most species have a single gene that encodes a centromeric histone variant while C. elegans has two: HCP-3 (also known as CeCENP-A) and CPAR-1. Before the replacement RNAi experiments showed that the HCP-3 is a functional isoform dominant, consistent with CPAR-1 does not become detectable in the embryo.

GFP :: CPAR-1 loaded onto meiotic chromosomes in diakinesis and enriched bivalents until meiosis I. Here we show that the CPAR-1 :: GFP signal loss of chromosome segregation precisely coincide with the homologous during anaphase I. The loss of GFP:: CPAR-1 signal reflects the proteolytic cleavage between GFP and histone fold of CPAR-1, as CPAR-1 :: GFP, where GFP fused to the C-terminus of CPAR-1, showed no GFP signal loss.

A focus of the candidates involved separase screen, proteases which initiate anaphase by splitting kleisin subunit of cohesin, in reaction to this division. Examination of the N-terminal tail sequence CPAR-1 expressing separase cleavage site and a mutation of the residues alleged signatures on this site removed cleavage reaction, as visualized by the retention of CPAR-1 :: GFP signal in separating homologous chromosomes in metaphase-anaphase transition of meiosis I.

Separase (phospho Ser801) Polyclonal Antibody
ABP50519-003ml	Abbkine	0.03ml	EUR 189.6
Description: A polyclonal antibody for detection of Separase phospho Ser801) from Human, Mouse. This Separase phospho Ser801) antibody is for WB , IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the phosphorylation site of S801

Separase (phospho Ser801) Polyclonal Antibody
ABP50519-01ml	Abbkine	0.1ml	EUR 346.8
Description: A polyclonal antibody for detection of Separase phospho Ser801) from Human, Mouse. This Separase phospho Ser801) antibody is for WB , IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the phosphorylation site of S801

Separase (phospho Ser801) Polyclonal Antibody
ABP50519-02ml	Abbkine	0.2ml	EUR 496.8
Description: A polyclonal antibody for detection of Separase phospho Ser801) from Human, Mouse. This Separase phospho Ser801) antibody is for WB , IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the phosphorylation site of S801

Separase (phospho Ser801) Polyclonal Antibody
ES1518-100ul	ELK Biotech	100ul	EUR 334.8
Description: A Rabbit Polyclonal antibody against Separase (phospho Ser801) from Human/Mouse. This antibody is tested and validated for WB, ELISA, IHC, IF, WB, ELISA

Separase (phospho Ser801) Polyclonal Antibody
ES1518-50ul	ELK Biotech	50ul	EUR 248.4
Description: A Rabbit Polyclonal antibody against Separase (phospho Ser801) from Human/Mouse. This antibody is tested and validated for WB, ELISA, IHC, IF, WB, ELISA

Separin Phospho-Ser801 (ESPL1 pS801) Antibody
20-abx326180	Abbexa	EUR 376.80 EUR 292.80	100 ug 50 ug

SEPARASE (Phospho-Ser801) Colorimetric Cell-Based ELISA Kit
EKC2000	BosterBio	100ul	EUR 686.4

Phospho-SEPARASE (Ser801) Colorimetric Cell-Based ELISA Kit (OKAG01525)
OKAG01525	Aviva Systems Biology	2 x 96 Wells	EUR 888
Description: Description of target: ;Species reactivity: Human: S801, Mouse: S1189;Application: ELISA;Assay info: Assay Type: Cell-Based Subtype: Phospho Detection Method: Colorimetric 450 nm;Sensitivity:

Neither whether or uncleavable CPAR-1 centromere-localized in mitosis and not locally around chromatin, indicating that activity has not been retained in the CPAR centromere-1. Although the function of CPAR-1 and cleavage depends separase that remains to be explained, these efforts reveal new substrate separase and provide in vivo biosensors to monitor activity at the beginning of meiosis I separase anaphase.